Method for microbial production of cyclic adenosine 3′, 5′-monophosphate

Abstract

The invention relates to a method for the production of adenosine 3′,5′-monophosphate (cAMP) by a permeable microbial strain which uses polyols for protecting activities of enzymes and a cAMP precursor and phosphate as substrates. Glucose is used as an energy provider and metal ions and an organic solvent such as acetone are used in the medium.

The invention claimed is: 
     
       1. A method for producing cyclic adenosine monophosphate by whole-cell biocatalysis, the method comprising:
 preparing a reaction solution comprising an enzyme activity protector, a substrate, an energy provider, a metal ion, and a permeable microbial strain and an organic solvent; and 
 conducting a whole-cell biocatalysis reaction in said reaction solution to obtain cyclic adenosine monophosphate, 
 wherein said enzyme activity protector is one or more polyhydric organic reagents selected from the group consisting of mannitol, maltitol, xylitol, lactitol, sorbitol, and glycol; said substrate comprises a cyclic adenosine monophosphate precursor and phosphate ion; and said energy provider is glucose; said metal ions are any one or more selected from magnesium ion and ammonium ion; said permeable microbial strain is any one strain selected from  arthrobacter , microbaterium and  corynebacterium ; and said organic solvent is acetone; 
 wherein the one or more polyhydric organic reagents is present in an initial reaction concentration of 0.001-10 g/L, the cyclic adenosine monophosphate precursor is present in an initial reaction concentration of 0.01-100 g/L, the phosphate on is present in an initial reaction concentration of 0.01-100 g/L, the glucose is present in an initial reaction concentration of 0.01-100 g/L, the metal ion is present in an initial reaction concentration of 0.001-10 g/L, the microbial strain is present in an amount of 10-1,000 g wet thallus/L, and the acetone is present in an initial reaction concentration of 0.01-50 mL/L. 
 
     
     
       2. The method for producing cyclic adenosine monophosphate by whole-cell biocatalysis according to  claim 1 , characterized in that said cyclic adenosine monophosphate precursor is adenine, adenosine, adenosine monophosphate, adenosine triphosphate, inosine, inosinic acid, or hypoxanthine. 
     
     
       3. The method for producing cyclic adenosine monophosphate by whole-cell biocatalysis according to  claim 1 , characterized in that the conducting a whole-cell biocatalysis reaction is conducted in aqueous solution for 2-200 hours under the condition of pH 4-10 and 25-38° C. 
     
     
       4. The method for producing cyclic adenosine monophosphate by whole-cell biocatalysis according to  claim 2 , characterized in that the conducting a whole-cell biocatalysis reaction is conducted in aqueous solution for 2-200 hours under the condition of pH 4-10 and 25-38° C. 
     
     
       5. The method for producing cyclic adenosine monophosphate by whole-cell biocatalysis according to  claim 1 , characterized in that said microbial strain is any one strain selected from  Brevibacterium liquefaciens, Arthrobacter roseoparaffincus, Escherichia coli, Bacillus subtilis , yeast and  Brevibacterium ammoniagenes.